Pyrosequencing is a sequencing by synthesis method that provides a quantitative measure of the proportion of bases at every position within any PCR-amplified sequence. We exploit this property of Pyrosequencing to obtain highly quantitative measures of DNA methylation in gene sequences as well as repetitive elements that allow for discriminating differences lower than 1% methylation.
Pyrosequencing is the gold standard for DNA methylation analysis in candidate sequences up to 100 bp in length. The laboratory is equipped with Pyrosequencing PSQ™HS 96 (Pyrosequencing Inc., Westborough, MA). Seven Biorad PCR units are used for pre-Pyrosequencing amplification.